title: Towards characterization of the glycoproteome of tomato (Solanum lycopersicum) fruit using Concanavalin A lectin affinity chromatography and LC-MALDI-MS/MS analysis.
abstract: The isolation and analysis of glycoproteins by coupling lectin affinity chromatography with MS has emerged as a powerful strategy to study the glycoproteome of mammalian cells. However, this approach has not been used extensively for the analysis of plant glycoproteins. As with all eukaryotes, N-glycosylation is a common post-translational modification for plant proteins traveling through the secretory pathway. Many such proteins are destined for the cell wall, or apoplast, where they play important roles in processes such as modifying cell wall structure, sugar metabolism, signaling, and defense against pathogens. Here, we describe a strategy to enrich for and identify secreted plant proteins based on affinity chromatography using the lectin Concanavalin A and two-dimensional liquid chromatography, together with matrix-assisted laser desorption/ionization MS analysis. The value of this approach is illustrated through the characterization of glycoproteins that are expressed in ripe tomato (Solanum lycopersicum) fruit, a developmental stage that is fundamentally linked with significant changes in cell wall structure and composition. This glycoprotein trap strategy allowed the isolation of a sub-proteome with an extremely high proportion of proteins that are predicted to be resident in the cell wall or secretory pathway, and the identification of new putative cell wall proteins.
authors.affiliation: Department of Plant Biology, Cornell University, Ithaca, NY, USA.
authors.forename: Carmen
authors.initials: C
authors.lastname: Catalá
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Howe
authors.forename: Simon
authors.initials: S
authors.lastname: Hucko
authors.forename: Jocelyn K C
authors.initials: JK
authors.lastname: Rose
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Thannhauser
mesh_terms: D002473:Cell Wall / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A / Q000737:chemistry; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D018551:Solanum lycopersicum / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006023:Glycoproteins; D010940:Plant Proteins; D020543:Proteome; D003208:Concanavalin A